Permount

Manufactured by Thermo Fisher Scientific

Sourced in United States, Canada, Japan, Germany, United Kingdom, Gabon, China

Permount is a mounting medium used in microscopy to permanently mount specimens on glass slides. It is a solvent-based, xylene-containing solution that dries to form a clear, resinous film, securing the specimen in place and providing optical clarity for microscopic examination.

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Lab products found in correlation

1 051 protocols using permount

Myelination and Histology Analysis of Spinal Cord

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Luxol fast blue staining was used to visualize myelin. Transverse SC cryostat sections were dehydrated by use of 85%, 95%, and 100% ethanol followed by xylene. They were incubated in 0.1% Luxol fast blue overnight at 56 °C (Thermo Fisher Scientific Inc.). The next day, the sections were differentiated in 0.05% lithium carbonate (Sigma Aldrich) and 70% ethanol, dehydrated with increasing concentrations of ethanol, and coverslipped using Permount (Thermo Fisher Scientific Inc.) mounting media. Pictures were obtained using a Nikon Eclipse 50i microscope equipped with a × 4 objective, using Ocular software (Digital Optics Ltd., Auckland, New Zealand).
Hematoxylin and eosin staining was performed with an automated stainer (Leica Autostainer XL, Nussloch, Germany). The SC sections were immersed in Harris hematoxylin and eosin stain. Following dehydration in 95% and 100% alcohol solutions, the section were coverslipped using Permount mounting media (Thermo Fisher Scientific Inc.) and pictures were taken with a Nikon Eclipse 50i microscope equipped with a × 10 objective and using the Ocular software (Digital Optics Ltd).

Mirabelli E., Ni L., Li L., Acioglu C., Heary R.F, & Elkabes S. (2019). Pathological pain processing in mouse models of multiple sclerosis and spinal cord injury: contribution of plasma membrane calcium ATPase 2 (PMCA2). Journal of Neuroinflammation, 16, 207.

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Quantifying Dendritic Spine Density in Hippocampal Neurons

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The protocol for spine density analysis was as previously described (Nishijima et al., 2014 (link)). Briefly, 4 weeks after the DZP administration, mice (N = 4 per group) were deeply anesthetized and intracardially perfused with 4% paraformaldehyde, and their brains were extracted and placed in PB. Coronal sections (250 μm) that included the hippocampus were prepared using a microslicer (VT-1000; Leica, Wetzlar, Germany). Neuronal spines were visualized using Lucifer yellow injection into pyramidal neurons of the CA1/3 by iontophoresis pump (BAB-600; Kation Scientific, Minneapolis, MN, United States) under a fluorescence microscope (BX51; Olympus, Tokyo, Japan). The sections were then mounted onto slides and coverslipped with Permount and SlowFade Gold Antifade Reagent (S36937; Invitrogen, Tokyo, Japan). Images of the Lucifer yellow-injected neurons were acquired using a laser confocal microscope (C1si; Nikon, Tokyo, Japan) with a 60 × oil-immersion lens. We then measured the density of spines on apical branches that were observed to be 100–200 μm from the cell body in the pyramidal cell layer of CA1 or CA3. Images (2–4 images/neuron) were acquired with zoom and were taken at 0.25 μm focal steps. Images were analyzed offline using NIS-Elements software (Nikon). The average number of spines per 20 μm of dendritic length was expressed as the spine density.

Furukawa T., Nikaido Y., Shimoyama S., Masuyama N., Notoya A, & Ueno S. (2021). Impaired Cognitive Function and Hippocampal Changes Following Chronic Diazepam Treatment in Middle-Aged Mice. Frontiers in Aging Neuroscience, 13, 777404.

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Immunohistochemical Analysis of CML and RAGE

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All specimens were formalin-fixed and paraffin-embedded. Deparaffinized tissue sections were rehydrated, and antigen retrieval performed by heating in a vegetable steamer in 10 mM citrate, pH 6.0 for 30mins. Endogenous peroxidase activity was blocked using 0.3% H₂O₂ in methanol for 30mins. Sections were washed and nonspecific binding was blocked using 2.5% horse serum and then incubated overnight at 4°C with target-specific primary antibody at a 1:100 dilution. CML primary antibody (ab27684) was from Abcam (Cambridge, MA). RAGE primary antibody (AB5601) was from Millipore (Billerica, MA). Sections were fixed in Permount (Invitrogen) and mounted on slides. All sections were examined using an Olympus BX50 microscope and pictures were taken using an Olympus DP 70 camera connected to DP Controller software (Olympus, Center Valley, PA). IHC scores were calculated using the formula: intensity X % positive tumor cells. Intensity staining was scored: 1 – weak, 2 – intermediate, 3 – strong and 4 - very strong. % positive tumor cells were scored: 1 – less than 10%, 2 – 10 to 30%, 3 – 30 to 60% and 4 – 60 to 100% positive cells. Fluorescence was quantified using image J.

Foster D., Spruill L., Walter K.R., Nogueira L.M., Fedarovich H., Turner R.Y., Ahmed M., Salley J.D., Ford M.E., Findlay V.J, & Turner D.P. (2014). AGE metabolites: A biomarker linked to cancer disparity?. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, 23(10), 2186-2191.

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Histological Examination of Liver Tissues

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Liver tissues were either formalin fixed and embedded in paraffin (FFPE), or embedded frozen in optimal cutting temperature (OCT) prior to cutting at either 5 µm or 8 µm, respectively, and then mounted onto charged glass slides. FFPE sections were processed in Citrisolv (Thermo Fisher Scientific, Waltham, MA) and rehydrated via incubation in graded ethanol concentrations. Sections were stained with hematoxylin and eosin (H&E), Sirius red, or Mason’s Trichrome, before mounting with Permount (Thermo Fisher, Waltham, MA).

Hudson S.V., Miller H.A., Mahlbacher G.E., Saforo D., Beverly L.J., Arteel G.E, & Frieboes H.B. (2019). Computational/experimental evaluation of liver metastasis post hepatic injury: interactions with macrophages and transitional ECM. Scientific Reports, 9, 15077.

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Histological Analysis of Reproductive Tissues

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Formalin-fixed, paraffin-embedded spleen, testes, or ovary tissue was sectioned into 5-μm-thick sagittal sections, placed on Superfrost microscope slides (Thermo Fisher Scientific, Hampton, NH, USA), and baked at 37°C overnight. The sections were de-paraffinized using two changes of xylene and rehydrated by immersing in 100%, 90%, and then 70% ethanol. The sections were stained for nuclear structures using Harris hematoxylin (Surgipath, Buffalo Grove, IL, USA) for 2 min, followed by differentiation in 1% acid alcohol (Surgipath) and treatment with Scott’s tap water for 2 min. Subsequently, the sections were counterstained for cytoplasmic structures using eosin (Surgipath) for 2 min. The slides were dehydrated with 70%, 90%, and 100% ethanol; cleared in xylene; and mounted using Permount (Thermo Fisher Scientific). Slides were imaged on a Nikon80i Upright microscope using NIS Elements BR software. Testis and ovary tissue histopathology was evaluated by the PennVet Comparative Pathology Core. This evaluation was performed in a blinded fashion.

Esquivel R.N., Patel A., Kudchodkar S.B., Park D.H., Stettler K., Beltramello M., Allen J.W., Mendoza J., Ramos S., Choi H., Borole P., Asija K., Bah M., Shaheen S., Chen J., Yan J., Durham A.C., Smith T.R., Broderick K., Guibinga G., Muthumani K., Corti D., Humeau L, & Weiner D.B. (2019). In Vivo Delivery of a DNA-Encoded Monoclonal Antibody Protects Non-human Primates against Zika Virus. Molecular Therapy, 27(5), 974-985.

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DCX Immunohistochemistry Protocol

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In the same manner, as described below, immunohistochemistry for DCX was performed (Park et al., 2019 (link)). The sections were treated with PBS for 10 min and then treated again with 1% H₂O₂ for 20 min. The sections were treated with goat anti-DCX antibody (1:1,000; Oncogene Research Product, Cambridge, UK) for 2 hr. The sections were treated with the biotinylated goat secondary antibody (1:500; Vector Laboratories, Burlingame, CA, USA) for another 1 hr, washed, and treated with ABC complex (1:100; Vector Laboratories). Using the 0.03% diaminobenzidine, labeling was visualized and the sections were mounted onto gelatin-coated slides. After air drying the slides at room temperature overnight, the coverslips were mounted using Permount (Thermo Fisher Scientific Inc., Waltham, MA, USA).

Ji E.S., Kim Y.M., Ko Y.J, & Baek S.S. (2020). Treadmill exercise in obese maternal rats during pregnancy improves short-term memory through neurogenesis in the hippocampus of rat pups. Journal of Exercise Rehabilitation, 16(5), 392-397.

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Histological Analysis of Tumor Sections

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Tissue was embedded in paraffin, sectioned at a thickness of 5 μm, and mounted onto positively-charged slides. Slides were baked in 65 °C for 1 h and paraffin was removed in xylene. Tissue was rehydrated in graded ethanol and rinsed with distilled water before staining with hematoxylin for 4 min. After thorough washing with tap water, slides were treated with bluing agent for 4 min and washed again in tap water for 1 min. Slides were rinsed in 95% ethanol and counterstained in eosin solution for 3 min. Slides were dehydrated through graded ethanol, cleared in xylenes, and cover-slipped with Permount (ThermoFisher Scientific, Houston, TX). Three representative sections from each tumor sonicated at the various PNP and control were evaluated qualitatively for hemorrhage and necrosis.

Aydin O., Lorsung R., Chandran P., Cohen G., Burks S.R, & Frank J.A. (2019). The proteomic effects of pulsed focused ultrasound on tumor microenvironments of murine melanoma and breast cancer models. Ultrasound in medicine & biology, 45(12), 3232-3245.

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Cresyl Violet Staining of Tissue Sections

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After wash with PBS, tissue sections were incubated in 0.1% cresyl violet (Sigma-Aldrich, St. Louis, MO) for 15 min. Stained sections were mounted with permount (Thermo Fisher scientific, Waltham, MA) after dehydration with a series of 70%, 95%, and 100% ethanol and xylene.

Horiuchi M., Smith L., Maezawa I, & Jin L.W. (2016). CX3CR1 ablation ameliorates motor and respiratory dysfunctions and improves survival of a Rett syndrome mouse model. Brain, behavior, and immunity, 60, 106-116.

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Visualizing Myelination and Cytochrome Oxidase

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One series of sections from each case was processed for myelin using the Gallyas^{25 (link)} silver procedure, which is based on the binding of colloidal silver to myelin for viewing by light microscopy. A second series of sections was processed for cytochrome oxidase^{26 (link)} to identify the original CO blobs. Stained sections were then mounted and dehydrated in an ascending series of ethanols (70% for 20 minutes, 95% for 10 minutes, 100% for 10 minutes), cleared in xylene, and coverslipped using Permount™ (Thermo Fisher Scientific, Waltham, MA, USA).

Rockoff E.C., Balaram P, & Kaas J.H. (2014). Patchy distributions of myelin and vesicular glutamate transporter 2 align with cytochrome oxidase blobs and interblobs in the superficial layers of the primary visual cortex. Eye and Brain, 6(Suppl 1), 19-27.

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Histological Liver Tissue Processing

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After macroscopic assessment (size, color, consistency), the resected liver specimens were fixated in 10% neutral buffered formalin for 12 h and subsequently dehydrated by passing the tissue through a series of increasing alcohol concentrations. After dehydration, the tissues were embedded in paraffin and trimmed to 3 µm sections. After deparaffinization and rehydration with decreasing strengths of alcohol, sections were stained with Mayer’s hematoxylin (Chroma, Münster) for 15 min, then washed in running tap water for 20 min followed by counterstaining with eosin (Chroma, Münster). After dehydration in 95% and absolute alcohols, sections were cleared in xylene (Merck, Darmstadt) and mounted in Permount (Thermo Fisher Scientific Inc).

Hillenbrand A., Kiebler B., Schwab C., Scheja L., Xu P., Henne-Bruns D., Wolf A.M, & Knippschild U. (2015). Prevalence of non-alcoholic fatty liver disease in four different weight related patient groups: association with small bowel length and risk factors. BMC Research Notes, 8, 290.

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